Download PDF
Thromb Haemost 2006; 95(05): 865-872
DOI: 10.1160/TH05-12-0786
Endothelium and Vascular Development
Schattauer GmbH

In vivo antithrombotic synergy of oral heparin and arginine: Endothelial thromboresistance without changes in coagulation parameters

Bruce Daniels
1   4221 South Western, Suite 4045, Oklahoma City, Oklahoma, USA
,
Robert J. Linhardt
2   Department of Chemistry and Chemical Biology, Biology and Chemical and Biological Engineering, Rensselaer Polytechnic Institute, Troy, New York, USA
,
Fuming Zhang
2   Department of Chemistry and Chemical Biology, Biology and Chemical and Biological Engineering, Rensselaer Polytechnic Institute, Troy, New York, USA
,
Wenjun Mao
3   Center for Marine Foods and Drugs, Qingdao Ocean University, Qingdao, China
,
Sandra M. Wice
4   Department of Veterinary Biomedical Sciences, University of Saskatchewan, Saskatoon, Saskatchewan, Canada
,
Linda M. Hiebert
4   Department of Veterinary Biomedical Sciences, University of Saskatchewan, Saskatoon, Saskatchewan, Canada
› Author AffiliationsFinancial support: This work was supported by Endomatrix Inc. Santa Rosa, California, a grant to Linda Hiebert from the Heart and Stroke Foundation of Saskatchewan and grants from the National Institutes of Health to Robert Linhardt (NIH HL52622 and GM38060).
Further Information

Publication History

Received 05 December 2005

Accepted after resubmission 10 March 2006

Publication Date:
01 December 2017 (online)

    Permissions and Reprints

Summary

On the basis of suggested clinical efficacy in an uncontrolled study in ninety-seven patients with unstable angina, an animal study was conducted to investigate antithrombotic synergy between orally administered heparin and arginine. A rat venous thrombosis model tested the difference in thrombus formation when heparin (7.5 mg/kg) and arginine (113 mg/kg) were administered, alone or in combination, by stomach tube with a minimum of 20 rats/group. Oral heparin, arginine, and heparin plus arginine reduced thrombus formation by 50%, 75%, and 90%, respectively,when compared to saline administration. Heparin was recovered from endothelium, yet there was little or no observable plasma anticoagulant activity. An orally administered lowmolecular-weight anticoagulant glycosaminoglycan mixture, sulodexide (7.5 mg/kg), showed an 88% reduction in stable thrombus formation when administered alone but showed no synergy with oral arginine. A 28-day study with oral sulodexide (2.9 mg/ kg) and arginine (43.9 mg/kg), 20 rats/group, showed antithrombotic activity with minimal anticoagulant activity indicating suitability for long term treatment. These findings suggest the endothelial localization of heparin and a synergistic antithrombotic effect for orally administered heparin and arginine.

Keywords

Heparin - thrombosis - endothelium - arginine - oral
 

  • References

  • 1 Jaques LB, Hiebert LM, Wice SM. Evidence from endothelium of gastric absorption of heparin and of dextran sulfates 8000. J Lab Clin Med 1991; 117: 122-30.
    PubMedGoogle Scholar
  • 2 Hiebert LM. Oral heparins. Clin Lab 2002; 48: 111-6.
    PubMedGoogle Scholar
  • 3 Hiebert LM, Wice SM, McDuffie NM. et al. The heparin target organ– the endothelium. Studies in a rat model. Quart J Med 1993; 86: 341-8.
    PubMedGoogle Scholar
  • 4 Hiebert LM, Ping T, Wice SM. Antithrombotic activity of orally administered low molecular weight heparin (logiparin) ina rat model. Haemostasis 2000; 30: 196-203.
    PubMedGoogle Scholar
  • 5 Napoli C, Ignarro LJ. Nitric oxide and atherosclerosis. Nitric Oxide 2001; 05: 88-97.
    CrossrefPubMedGoogle Scholar
  • 6 Armstead VE, Minchenko AG, Schuhl RA. et al. Regulation of P-selectin expression in human endothelial cells by nitric oxide. Am J Physiol 1997; 273: 740-6.
    PubMedGoogle Scholar
  • 7 Ignarro LJ, Cirino G, Casini A. et al. Nitric oxide as a signaling molecule in the vascular system: an overview. J Cardiovasc Pharmacol 1999; 34: 879-86.
    CrossrefPubMedGoogle Scholar
  • 8 Keha D, Thieme A, Kurer I. et al. Inactivation of platelet glycoprotein IIb/IIIa receptor by nitric oxide donor 3-morpholino-sydnonimine. Blood Coagul Fibrinolysis 2003; 14: 327-34.
    CrossrefPubMedGoogle Scholar
  • 9 Stief TW, Weippert M, Kretschmer V. et al. Arginine inhibits hemostasis activation. Thromb Res 2001; 104: 265-74.
    CrossrefPubMedGoogle Scholar
  • 10 Hileman R, Fromm J, Wieler J. et al. Glycosaminoglycan-protein interations:definition of consensus sites in glycosaminoglycan binding proteins. Bioessays 1998; 20: 156-67.
    CrossrefPubMedGoogle Scholar
  • 11 Fromm JR, Hileman RE, Caldwell EE. et al. Differences in the interaction of heparin with arginine and lysine and the importance of these basic amino acids in the binding of heparin to acidic fibroblast growth factor. Arch Biochem Biophys 1995; 323: 279-87.
    CrossrefPubMedGoogle Scholar
  • 12 Florian JA, Kosky JR, Ainslie K. et al. Heparan sulfate proteoglycan is a mechanosensor on endothelial cells. Circ Res 2003; 93: 136-42.
    CrossrefPubMedGoogle Scholar
  • 13 Pinhal MA, Walenga JM, Jeske W. et al. Antithrombotic agents stimulate the synthesis and modify the sulfation pattern of a heparan sulfate proteoglycan from endothelial cells. Thromb Res 1994; 74: 143-53.
    CrossrefPubMedGoogle Scholar
  • 14 Irokawa M, Nishinaga M, Ikeda U. et al. Endothelial-derived nitric oxide preserves anticoagulant heparan sulfate expression in cultured porcine aortic endothelial cells. Atherosclerosis 1997; 135: 9-17.
    CrossrefPubMedGoogle Scholar
  • 15 Lloyd-Jones DM, Bloch KD. The vascular biology of nitric oxide and its role in atherogenesis. Annu Rev Med 1996; 47: 365-75.
    CrossrefPubMedGoogle Scholar
  • 16 Stevens RL, Colombo M, Gonzales JJ. et al. The glycosaminoglycans of the human artery and their changes in atherosclerosis. J Clin Invest 1976; 58: 470-81.
    CrossrefPubMedGoogle Scholar
  • 17 Bingley JA, Hayward IP, Girjes AA. et al. Expression of heparan sulphate N-deacetylase/N-sulphotransferase by vascular smooth muscle cells. HistochemJ 2002; 34: 131-7.
    PubMedGoogle Scholar
  • 18 Bombeli T, Karsan A, Tait JF. et al. Apoptotic vascular endothelial cells become procoagulant. Blood 1997; 89: 2429-42.
    PubMedGoogle Scholar
  • 19 Constantinescu AA, Vink H, Spaan JA. Endothelial cell glycocalyx modulates immobilization of leukocytes at the endothelial surface. Arterioscler Thromb Vasc Biol 2003; 23: 1541-7.
    CrossrefPubMedGoogle Scholar
  • 20 Ding K, Sandgren S, Mani K. et al. Modulations of glypican-1 heparan sulfate structure by inhibition of endogenous polyamine synthesis. Mapping of spermine-binding sites and heparanase, heparin lyase, and nitric oxide/nitrite cleavage sites. J Biol Chem 2001; 276: 46779-91.
    PubMedGoogle Scholar
  • 21 Ruberg FL, Leopold JA, Loscalzo J. Atherothrombosis: plaque instability and thrombogenesis. Prog Cardiovasc Dis 2002; 44: 381-94.
    CrossrefPubMedGoogle Scholar
  • 22 Ofosu FA. Pharmacological actions of sulodexide. Semin Thromb Hemost 1998; 24: 127-38.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 23 Wallentin L, Lagerqvist B, Husted S. et al. Outcome at1 year after an invasive compared with a noninvasive strategy in unstable coronary-artery disease: the FRISC II invasive randomised trial. FRISC II Investigators. Fast Revascularisation during Instability in Coronary artery disease. Lancet 2000; 356: 9-16.
    CrossrefPubMedGoogle Scholar
  • 24 Blake OR, Ashwin JG, Jaques LB. An assay for the antithrombotic activity of anticoagulants. J Clin Path 1959; 12: 118-22.
    CrossrefPubMedGoogle Scholar
  • 25 Hiebert LM, Jaques LB. Heparin uptake on endothelium. Artery 1976; 02: 26-37.
    PubMedGoogle Scholar
  • 26 Hiebert LM, Wice SM, Ping T. et al. Tissue distribution and antithrombotic activity of unlabeled or 14C-labeled porcine intestinal mucosal heparin following administration to rats by the oral route. Can J Physiol Pharm 2000; 78: 307-20.
    CrossrefPubMedGoogle Scholar
  • 27 Hiebert LM, Wice SM, Ping T. et al. Antithrombotic efficacy ina rat model of the low molecular weight heparin, reviparin sodium, administered by the oral route. Thromb Haemost 2001; 85: 114-8.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 28 Engelberg H. Orally ingested heparin is absorbed in humans. Clin Appl Thromb Hemost 1995; 01: 283-5.
    CrossrefPubMedGoogle Scholar
  • 29 Hiebert LM, Wice SM, Jaques LB. et al. Orally administered dextran sulfate is absorbed in HIV-positive individuals. J Lab Clin Med 1999; 133: 161-70.
    CrossrefPubMedGoogle Scholar
  • 30 Hiebert LM, Wice SM, Ping T. Increased plasma anti-Xa activity and recovery of heparin from urine suggest absorption of orally administered unfractionated heparin in human subjects. J Lab Clin Med 2005; 145: 151-5.
    CrossrefPubMedGoogle Scholar
  • 31 Pinel C, Wice SM, Hiebert LM. Orally administered heparins prevent arterial thrombosis in a rat model. Thromb Haemost 2004; 91: 919-26.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 32 Kouretas PC, Hannan RL, Kapur NK. et al. Nonanticoagulant heparin increases endothelial nitric oxide synthase activity: Role of inhibitory guanine nucleotide proteins. J Mol Cell Cardiol 1998; 30: 2669-82.
    CrossrefPubMedGoogle Scholar
  • 33 Florian JA, Kosky JR, Ainslie K. et al. Heparan sulfate proteoglycan is a mechanosensor on endothelial cells. Circ Res 2003; 93: 136-42.
    CrossrefPubMedGoogle Scholar
  • 34 Li Y, Wang WP, Wang HY. et al. Intragastric administration of heparin enhances gastric ulcer healing through a nitric oxide-dependent mechanism in rats. Eur J Pharmacol 2000; 399: 205-14.
    CrossrefPubMedGoogle Scholar
  • 35 Hoffmann J, Haendeler J, Aicher A. et al. Aging enhances the sensitivity of endothelial cells toward apoptotic stimuli: important role of nitric oxide. Circ Res 2001; 89: 709-15.
    CrossrefPubMedGoogle Scholar
  • 36 Murata K, Murata A, Yoshida K. Heparan sulfate isomers in cerebral arteries of Japanese women with aging and with atherosclerosis--heparitinase and highperformance liquid chromatography determinations. Atherosclerosis 1997; 132: 9-17.
    CrossrefPubMedGoogle Scholar
  • 37 Kolodgie FD, Burke AP, Farb A. et al. Differential accumulation of proteoglycans and hyaluronan in culprit lesions: insights into plaque erosion. Arterioscler Thromb Vasc Biol 2002; 22: 1642-8.
    CrossrefPubMedGoogle Scholar